QIAamp RNA Blood Mini Kit

从新鲜的全血样本中纯化胞内RNA

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QIAamp RNA Blood Mini Kit (50)

Cat. No. / ID: 52304

For 50 RNA preps: 50 QIAamp Mini Spin Columns, 50 QIAshredder Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free reagents and buffers

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KitBuffer

QIAamp RNA Blood Mini Kit

Buffer AW1

QIAamp RNA Blood Mini Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的（再）订购

特点

快速纯化高品质的即用型RNA
无需有机提取或乙醇沉淀
重复性好，产量高
完全去除污染物和抑制剂

产品详情

QIAamp RNA Blood Mini Kit使用硅胶膜，可从多至1.5 ml的人类新鲜全血中纯化胞内RNA。可以使用常规的抗凝剂如柠檬酸盐、肝素或EDTA等稳定血液样本。使用QIAshredder离心柱对样本进行均化处理后进行快速离心，从而简化了纯化RNA的流程。可在QIAcube全自动核酸纯化仪上全自动完成纯化处理。

绩效

QIAamp纯化流程可完全去除RNases、污染物和酶抑制剂，制备的高品质RNA适用于多种下游应用（参见" High-quality RNA for northern analysis"和" Reliable RT-PCR analysis"）。

QIAamp RNA Blood Mini Kit制备的RNA品质高，含有最少量的DNA。RNA的纯化不可避免存在DNA污染。但对于某些痕量DNA敏感的RNA应用则必须去除残留的DNA，对于这些应用，QIAGEN RNase-Free DNase Set（可选购）为QIAamp RNA纯化流程提供简便的柱上DNase酶处理操作，去除残留DNA。

查看图表

High-quality RNA for northern analysis.

Reliable RT-PCR analysis.

原理

QIAamp RNA Blood Mini Kit使用硅胶膜技术纯化细胞RNA，无需苯酚-氯仿抽提。RNA特异性结合到QIAamp硅胶膜上，污染物流出。通过两步高效洗涤完全去除二价阳离子和蛋白质等PCR抑制剂，再用水或试剂盒中的缓冲液洗脱结合在离心柱上的纯DNA。

QIAamp技术从新鲜的全血样本或其他样本来源中纯化的总细胞RNA，可即时用于PCR和印迹实验中。QIAamp样本制备技术完全得到验证认可。

程序

QIAamp RNA Blood Mini Kit使用快速离心操作简化从血液中纯化RNA的操作流程（参见""）。选择性裂解红细胞，离心收集白细胞。在高度变性条件下白细胞裂解，RNases很快失活。使用QIAshredder离心柱均质化后，将样本上样到QIAamp离心柱。总RNA结合到QIAamp膜，污染物流出，在30–100 µl的无RNase水中洗脱残留的纯RNA（试剂盒提供），可即时用于多种下游应用。

查看图表

QIAamp RNA Blood Mini procedure.

应用

QIAamp RNA Blood Mini Kit可从多至1.5 ml的新鲜人体全血中纯化细胞RNA。可以使用常规的抗凝剂如柠檬酸盐、肝素或EDTA等稳定血液样本。也可从组织样本中纯化得到总细胞RNA。

辅助数据和图表

QIAamp RNA Blood Mini procedure.

Specifications

Features	Specifications
ApplicationsZH	PCR, real-time PCR, microarray
Elution volume	30–100 µl
Main sample type	Whole blood, tissue
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Cellular RNA
Sample amount	50–1.5 ml
Format	Spin columns
Processing	Manual (centrifugation)
Time per run or per prep	<1 hour
Technology	Silica technology
Yield	1–5 µg

资源

For total RNA purification from human whole blood

Poster for download

Download Safety Data Sheets for QIAGEN product components.

FAQ

One milliliter of healthy human blood consists of cell types in approximately the following numbers:

Leucocytes (function: immune response) 4–7 x 106 cells
Thrombocytes (function: wound closing) 3–4 x 108 cells
Erythrocytes (function O2 and CO2 transport) 5 x 109 cells

FAQ ID -2951

Several kit options are available for this application. We recommend using the PAXgene Blood RNA System, which enables the collection, stabilization and transportation of 2.5 ml human whole blood samples, and subsequent rapid and efficient isolation of cellular RNA.

Other products for the isolation of RNA from whole human blood are the QIAamp RNA Blood Mini Kit and the RNeasy Midi Kit for processing up to 1.5 ml and 10 ml human whole blood, respectively.

FAQ ID -304

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

For more information on activity of QIAGEN Protease and Proteinase K in various buffers please click here.

FAQ ID -761

Yes. The RNeasy MinElute Cleanup Kit has been developed specifically for cleaning up and concentrating RNA samples. You can also follow the protocols for RNA cleanup in the RNeasy Mini and RNeasy Midi/Maxi Handbook.

FAQ ID -286

Yes. Even though buffer RDD in the RNase-Free DNase Set is optimized for on-column DNase digestion, the buffer is also well-suited for efficient DNase digestion in solution. Please note that the reaction must be cleaned up after the off-column DNase digest to remove the enzyme and buffer RDD, which will interfere with subsequent RT reactions.

A protocol for in-solution DNase digestion using the RNase-Free DNase Set can be found in Appendix C of the RNeasy MinElute Cleanup Handbook. For subsequent RNA Cleanup, use either the RNeasy MinElute Cleanup Kit, or follow the instructions for RNA Cleanup in the RNeasy Mini Handbook.

FAQ ID -619

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.

For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.

FAQ ID -635

For RNA isolated on the BioRobot EZ1 and BioRobot M48:

The RNA can be directly applied to the Agilent Bioanalyzer, since it is being denatured in the final protocol steps of these isolation procedures.

For RNA prepared with all other QIAGEN RNA Isolation Products:

We recommend to denature the samples in a water bath for 2 min at 70°C, and then place them directly on ice prior to loading them onto the Agilent Bioanalyzer.

FAQ ID -528

The A₂₆₀/ A₂₈₀ ratio is influenced considerably by pH. Since water is not buffered, the pH and the resulting A₂₆₀/A₂₈₀ ratio can vary greatly. Lower pH results in a lower A₂₆₀/ A₂₈₀ ratio and a reduced sensitivity to protein contamination*.

For accurate values, we recommend measuring absorbance in 10 mM Tris·Cl, pH 7.5. Always be sure to calibrate the spectrophotometer with the same solution. Please see the Appendix sections in the RNeasy handbooks for additional information.

* Wilfinger, W.W., Mackey, M., and Chomczynski, P. (1997) Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques 22, 474.

FAQ ID -97

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12

The concentration of RNA isolated with RNeasy Kits can be determined by measuring the absorbance at 260 nm (A260) in a spectrophotometer. Absorbance readings should be greater than 0.15 to ensure significance. An absorbance of 1 unit at 260 nm corresponds to 40 µg of RNA per ml (A260 = 1 = 40 µg/ml). This relationship is valid for measurements in water. Therefore, dilute RNA in water to quantify it spectrophotometrically.

An example of the calculations involved in RNA quantification is shown below. Use the buffer in which the RNA is diluted to zero the spectrophotometer:

Volume of RNA sample = 100 µl
Dilution = 10 µl of RNA sample + 490 µl distilled water (1/50 dilution)
Absorbance of diluted sample measured in a 1 ml cuvette (RNase-free): A260 = 0.23
Concentration of original RNA sample = 40 x A260 x dilution factor = 40 x 0.23 x 50
RNA concentration: 460 µg/ml
Total yield = concentration x volume of sample (ml) = 460 µg/ml x 0.1 ml
RNA yield: 46 µg

For additional information on RNA quantitation and handling, see the Appendix section in the RNeasy Mini Handbook.

FAQ ID -32